Transgenic mice containing alpha-endosulfine gene disruptions

Abstract

The present invention relates to transgenic animals, as well as compositions and methods relating to the characterization of gene function. Specifically, the present invention provides transgenic mice comprising mutations in a alpha-endosulfine gene. Such transgenic mice are useful as models for disease and for identifying agents that modulate gene expression and gene function, and as potential treatments for various disease states and disease conditions. The present invention also relates to diabetes and diabetic condition, as it demonstrates the role of the alpha-endosulfine in diabetes and diabetic conditions. The present invention further relates to weight gain and weight related conditions, such as obesity, and demonstrates the role of the alpha-endosulfine in weight gain and weight related conditions, such as obesity. In accordance with these aspects, the present invention provides methods and compositions useful in identifying, testing, and providing treatments for diabetes and diabetic conditions, weight gain and weight related conditions such as obesity.

We claim:  
     
         1 . A targeting construct comprising: 
 (a) a first polynucleotide sequence homologous to an alpha-endosulfine gene;    (b) a second polynucleotide sequence homologous to the alpha-endosulfine gene; and    (c) a selectable marker.    
     
     
         2 . The targeting construct of  claim 1 , wherein the targeting construct further comprises a screening marker.  
     
     
         3 . A method of producing a targeting construct, the method comprising: 
 (a) providing a first polynucleotide sequence homologous to an alpha-endosulfine gene;    (b) providing a second polynucleotide sequence homologous to the alpha-endosulfine;    (c) providing a selectable marker; and    (d) inserting the first sequence, second sequence, and selectable marker into a vector, to produce the targeting construct.    
     
     
         4 . A method of producing a targeting construct, the method comprising: 
 (a) providing a polynucleotide comprising a first sequence homologous to a first region of the alpha-endosulfine gene and a second sequence homologous to the alpha-endosulfine gene; and    (b) inserting a positive selection marker in between the first and second sequences to form the targeting construct.    
     
     
         5 . A cell comprising a disruption in an alpha-endosulfine gene.  
     
     
         6 . The cell of  claim 5 , wherein the cell is a murine cell.  
     
     
         7 . The cell of  claim 6 , wherein the murine cell is an embryonic stem cell.  
     
     
         8 . A non-human transgenic animal comprising a disruption in an alpha-endosulfine gene.  
     
     
         9 . A cell derived from the non-human transgenic animal of  claim 8 .  
     
     
         10 . A method of producing a transgenic mouse comprising a disruption in the alpha-endosulfine gene, the method comprising: 
 (a) introducing the targeting construct of  claim 1  into a cell;    (b) introducing the cell into a blastocyst;    (c) implanting the resulting blastocyst into a pseudopregnant mouse, and    (d) identifying the transgenic mouse comprising a disruption in the alpha-endosulfine gene.    
     
     
         11 . A method of identifying an agent that modulates the expression or function of alpha-endosulfine, the method comprising: 
 (a) providing a non-human transgenic animal comprising a disruption in an alpha-endosulfine gene;    (b) administering an agent to the non-human transgenic animal; and    (c) determining whether the expression or function of alpha-endosulfine in the non-human transgenic animal is modulated.    
     
     
         12 . A method of identifying an agent that modulates the expression or function of alpha-endosulfine, the method comprising: 
 (a) providing a cell comprising a disruption in an alpha-endosulfine gene;    (b) contacting the cell with an agent; and    (c) determining whether expression or function of alpha-endosulfine is modulated.    
     
     
         13 . An agent identified by the method of  claim 11  and  claim 12 .  
     
     
         14 . The non-human transgenic animal of  claim 8 , wherein the transgenic animal exhibits less body weight than wild-type mice  
     
     
         15 . The non-human transgenic animal of  claim 8 , wherein the transgenic animal exhibits improved glucose tolerance.  
     
     
         16 . The non-human transgenic animal of  claim 8 , wherein the transgenic animal exhibits blood glucose sensitivity to exogenous insulin.  
     
     
         17 . The non-human transgenic animal of  claim 8 , wherein the transgenic animal exhibits impaired glucose secretion while on a high fat diet.  
     
     
         18 . The non-human transgenic animal of  claim 8 , wherein the transgenic animal exhibits hyperactivity.  
     
     
         19 . A method of screening for biologically active agents, the method comprising: 
 (a) combining a putative agent with a mammalian alpha-endosulfine polypeptide; and    (b) detecting an effect of said agent on alpha-endosulfine activity; wherein detection of a decrease or an increase in alpha-endosulfine activity is indicative of a biologically active agent.    
     
     
         20 . A method of screening for biologically active agents, the method comprising: 
 (a) combining a putative agent with an isolated cell comprising a nucleic acid encoding a mammalian alpha-endosulfine gene or a alpha-endosulfine promoter sequence operably linked to a reporter gene; and    (b) detecting an effect of said agent on alpha-endosulfine activity; wherein detection of a decrease or an increase in alpha-endosulfine activity is indicative of a biologically active agent